Metabolic diseases such as obesity, type 2 diabetes, metabolic dysfunction-associated steatotic liver disease, have become major global public health challenges. Their pathogenesis is closely linked to dysregulation of immune and inflammatory responses. This review summarizes the central role of immunoinflammation underlying metabolic diseases, with emphasis on the interactive mechanisms between the immune system and metabolic system. It is broadly appreciated that metabolic disturbances trigger chronic low-grade inflammation, which exacerbates insulin resistance, adipose tissue dysfunction, and multi-organ damage through the involvement of both innate and adaptive immune cells. Among these, metabolic reprogramming of macrophages, the dual role of neutrophil extracellular traps in vascular injury and fibrosis, and the regulatory functions of Tregs represent critical nodes in diseases progression. From this metaflammation perspective, intervention strategies are proposed at multiple levels: Lifestyle modifications serve as a foundation to effectively improve immune-inflammatory status; exploring the immunomodulatory potential of existing drugs for repurposing in metabolic disease treatment, such as lansoprazole; and emerging therapies, such as GLP-1 receptor agonists and 11β-HSD1 inhibitors also demonstrate strong effect on alleviating inflammatory responses. Collectively, these important advances not only deepen our understanding of the immunological mechanisms underlying metabolic diseases, but also lay a theoretical foundation for developing novel strategies for the prevention and treatment of metabolic diseases, highlighting promising potential in extending healthspan.

Innate lymphoid cells (ILCs) mainly consist of natural killer (NK) cells, type 1 ILC (ILC1), type 2 ILC (ILC2), and type 3 ILC (ILC3), all exhibiting rapid responsiveness to pathogens and tumor cells. NK cells execute antitumor and antiviral functions through cytotoxic activity, cytokine secretion, and antibody-dependent cell-mediated cytotoxicity (ADCC), whereas ILC1, ILC2, and ILC3 modulate immune regulation and inflammatory responses within tissue microenvironments. Emerging evidence highlights the pivotal role of metabolic reprogramming in the development, activation, and effector functions of ILCs. Recent studies have further revealed that metabolic pathways are the core link regulating the development, maturation, activation and functional execution of ILCs. By precisely regulating the metabolic flux of substances such as glucose, amino acids and lipids, ILCs can achieve functional transition from a quiescent state to an effector state, while metabolic imbalance directly leads to abnormalities in their immune effects. This review systematically summarizes the metabolic characteristics of NK cells and other ILC subsets, and also discusses their underlying molecular mechanisms. Furthermore, we discuss how metabolic dysregulation in NK cells and ILCs contributes to pathological conditions such as obesity, cancer, and chronic viral infections, alongside potential therapeutic interventions. Finally, we elaborate on novel immunotherapeutic strategies targeting the metabolic reprogramming of ILCs, addressing unresolved questions in this rapidly evolving field. These insights establish a theoretical framework for developing next-generation ILC-based immunotherapies.

Aortic aneurysm (AA), an irreversible pathological dilation of the aorta exceeding 50% of its normal diameter, significantly increases the risk of aortic dissection and rupture-related sudden death, causing approximately 200 000 global deaths annually. AA can occur in various aortic segments and is primarily categorized into thoracic aortic aneurysm (TAA) and abdominal aortic aneurysm. TAA, though less common, has a worse prognosis. While the etiology and molecular mechanisms differ between the two types, both are closely linked to inflammatory responses and the activation of various proteases. This review will focus on the general cellular and molecular events applicable to both conditions. AA is usually asymptomatic, and even when detected via ultrasonography, it is usually monitored due to not meeting surgical criteria. As no effective medication currently exists to halt its growth or reduce rupture risk, this review will briefly cover animal and clinical drug trials to explore potential therapeutic strategies.

Chemotherapy-induced immune-priming cell death (ICD) is a unique form of cell death. By inducing this type of cell death through specific methods, cells can express damage-associated molecular patterns (DAMPs). These DAMPs can act as antigens to activate the host's immune system, thereby enhancing antitumor immune responses. Its clinical application has also attracted widespread attention. This review summarizes the molecular mechanisms of ICD, including pathways such as endoplasmic reticulum stress, autophagy, and necroptosis, as well as the mechanisms by which various chemotherapeutic drugs induce ICD. Additionally, this review explores the application of ICD in clinical practice, discusses its potential to provide new ideas and challenges for cancer treatment by categorizing it according to different types of cancer, and examines the possibility of applying ICD mechanisms in other diseases.

Multiple myeloma (MM) is a malignant hematological cancer that originates from plasma cells in the bone marrow. The disease is characterized by abnormal proliferation of clonal plasma cells and excessive secretion of monoclonal immunoglobulin. High protein load is a common feature of various malignant tumors, resulting from abnormal protein synthesis, folding errors, post-translational modifications (PTMs), and imbalances in the protein degradation system. The bone marrow microenvironment also plays a role in these processes. Interestingly, while high protein levels are toxic to normal cells, cancer cells actually rely on this stress to grow and survive. MM cells, in particular, produce large amounts of immunoglobulins, which contribute to their malignant progression. This review provides a comprehensive overview of current and investigational treatments for MM, as well as the key molecular pathways involved in maintaining protein homeostasis. Special attention is given to the role of PTMs in shaping the high protein load phenotype in MM cells. These findings are in line with the traditional Chinese medicine concept of "cancer toxin" and may offer new insights for developing therapeutic strategies that target proteotoxic stress.

Transcription factors (TFs) constitute a class of proteins that regulate gene transcription through sequence-specific DNA binding and thereby govern a broad spectrum of cellular processes and functions. Recent rapid advances in structural pharmacology have substantially enhanced mechanistic understanding of TF conformational transitions, ligand recognition features, and interaction networks with coregulatory partners, establishing a critical structural framework for rational targeted drug design. Nuclear receptors represent a structurally conserved yet functionally diverse family of ligand-regulated TFs that are extensively involved in major physiological and pathological processes, including development, metabolism, immunity, inflammation, and tumorigenesis. This review systematically summarizes the domain organization of prototypical nuclear receptors and the conformational characteristics of their ligand-binding domains, and highlights recent progress in nuclear receptor structural pharmacology. It further examines the structural basis and therapeutic potential of "nuclear receptor-like" family proteins as emerging drug targets, with representative examples drawn from hypoxia-inducible factors (HIF) and the aryl hydrocarbon receptor (AHR). In addition, advances in structure-guided discovery of compounds targeting other TF systems, including STAT3, p53/MDM2, and MYC/MAX, are discussed. Finally, the review provides a forward-looking perspective on the application of emerging technologies, such as AI-assisted structure prediction and dynamic conformational simulation, in TF-targeted drug discovery.

Abnormal aggregates with degradation-resistant properties are the core pathological hallmark of neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), Parkinson's disease (PD), and Huntington's disease (HD). Emerging studies have revealed that the pathogenesis of these diseases is closely linked to abnormalities in biomolecular condensates within cells. This review aims to systematically outline the relevant physiological and pathological functions of biomolecular condensates, summarize the regulatory mechanisms underlying phase separation of disease-related proteins and their associations with biomolecular condensates under physiological and pathological conditions, reveal the roles of these proteins in mediating neuronal dysfunction and death, and finally summarize the current therapeutic strategies targeting biomolecular condensates for intervention in neurodegenerative diseases.

Pulmonary fibrosis (PF) is a chronic interstitial lung disease with an average post-diagnosis survival of only 3-5 years. It is commonly characterized by dry cough and progressive dyspnea, ultimately leading to respiratory failure due to loss of gas exchange function. However, its exact pathogenic triggers and mechanisms remain unclear, and effective therapeutic drugs and approaches are still lacking. Therefore, further exploration of the pathogenesis of pulmonary fibrosis and the identification of new therapeutic targets are of great significance for its treatment and prognosis. Annexin A3 (ANXA3), a member of the calcium-dependent phospholipid-binding protein family, plays important roles in various pathophysiological processes such as membrane trafficking, inflammatory responses, and tumor progression. Nevertheless, its function and mechanism in pulmonary fibrosis have not yet been elucidated. In this study, using both in vivo animal models and in vitro cell experiments, we investigated the expression changes and functional mechanisms of ANXA3 in pulmonary fibrosis. In a bleomycin (BLM)-induced mouse model of pulmonary fibrosis, Western blot, quantitative real-time PCR, and immunofluorescence staining revealed that both the protein and mRNA levels of ANXA3 were significantly downregulated. To further clarify the role of ANXA3, a series of experiments were conducted using human embryonic lung fibroblasts (MRC-5). The results showed that knockdown of ANXA3 promoted the upregulation of fibrosis-related genes and significantly increased the expression of the extracellular matrix protein FN1. Meanwhile, EdU proliferation assays and wound healing assays demonstrated that ANXA3 knockdown markedly enhanced the proliferation and migration abilities of MRC-5 cells. In contrast, overexpression of ANXA3 significantly suppressed TGF-β1-induced fibrotic responses, including reduced expression of fibrotic markers, decreased extracellular matrix deposition, and weakened cell proliferation and migration. This study reveals that ANXA3 is downregulated during pulmonary fibrosis progression and plays a key inhibitory role in fibroblast activation and fibrogenesis, suggesting that ANXA3 may serve as an endogenous anti-fibrotic regulatory factor. These findings not only enrich the understanding of the ANXA protein family in the regulatory network of pulmonary fibrosis, but also provide experimental evidence for the future development of ANXA3-targeted therapeutic strategies for pulmonary fibrosis. This experiment was approved by the Ethics Committee of the School of Pharmacy, Harbin Medical University (approval number: IRB3035621).

Traumatic brain injury (TBI) is a common neurological disorder characterized by high incidence, disability, and mortality rates, posing a substantial global health burden and socioeconomic burden. The complexity of its pathology and the scarcity of effective clinical treatments present significant challenges for drug development. Various animal and in vitro models have been established to simulate TBI and its pathological sequelae, serving as essential tools for mechanistic exploration and drug discovery. This article provides a systematic review of the non-clinical evaluation framework for TBI, including the establishment of common models and key behavioral and molecular pharmacodynamic endpoints. Furthermore, it proposes several recommendations for standardizing the evaluation strategies of anti-TBI drugs, aiming to offer a structured reference for future research in this field.

Gouty arthritis is an inflammatory joint disease triggered by urate crystal deposition, with nucleotide binding oligomerization domain (NOD)-like receptor protein 3 (NLRP3) inflammasome overactivation serving as the core driver of its acute inflammatory response. Although NLRP3-targeting drugs have advanced to clinical trials, their clinical application remains limited by adverse effects and uncertain long-term risks. In this context, traditional Chinese medicine (TCM), with its multi-component, multi-target, and synergistic regulatory properties, offers novel strategies for the prevention and treatment of gout. Studies have shown that TCM exerts multidimensional therapeutic effects, including alleviation of acute joint inflammation, reduction of serum uric acid levels, and prevention of disease recurrence, underscoring its potential for systemic intervention. This review systematically summarizes the mechanisms by which active components and formulations of TCM modulate key aspects of NLRP3 inflammasome activation pathways. These include inhibition of upstream initiation signals, blockade of activation signal transduction, direct interference with inflammasome assembly, and enhancement of autophagic clearance, collectively leading to suppressed release of key inflammatory mediators such as IL-1β. This review aims to provide theoretical foundations and directional insights for further research and clinical application of TCM regulating NLRP3 in gouty arthritis treatment.

The central nervous system (CNS) serves as the core for regulating human body functions. The structural and metabolic networks of brain regions exhibit significant heterogeneity, with distinct brain regions having precisely divided metabolic characteristics. For CNS diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), and ischemic stroke (IS), their pathological mechanisms are often accompanied by metabolic disorders in specific brain regions. Notably, such metabolic perturbations precede macroscopic structural damage, thus providing a promising avenue for the identification of biomarkers for the early diagnosis of these disorders. However, traditional diagnostic methods and therapeutic effect assessment approaches have limitations. Specifically, they fall short in identifying subtle abnormalities in early metabolic levels, characterizing the spatial distribution of metabolites, and accurately determining whether treatments can reverse the metabolism-related pathological disorders. Based on mass spectrometry imaging technology, spatial metabolomics enables in situ detection and spatial localization of hundreds of metabolites in CNS tissue sections at a spatial resolution of the micrometer level. It not only fills the gap in technologies for detecting early metabolic abnormalities in diseases but also drives the research on disease pathological mechanisms from the "macroscopic structure" to the "molecular spatial network" level. Consequently, it provides crucial technical support for the screening of disease biomarkers, the analysis of disease mechanisms, and the evaluation of drug therapeutic effects. This article systematically reviews the main technologies of spatial metabolomics and deeply explores its current application status in CNS disease research. The aim of this review is to offer theoretical support and practical references for advancing the development of metabolomics in this field.

Antibody drug conjugates (ADCs) are a novel class of cancer therapeutics, structurally composed of monoclonal antibodies conjugated to cytotoxic agents via linkers. They offer advantages such as good tolerability, precise target recognition, and minimal impact on non-cancerous cells. To date, 19 ADCs have been approved for marketing, and over 300 ADCs are in various clinical trial phases. This review systematically outlines the developmental history of the three core components of ADCs—antibodies, payloads, and linkers. It aims to establish a foundational knowledge framework in ADCs development, thereby promoting technological dissemination and advancing awareness in the field. We anticipate that this review will serve as a foundational stepping stone for researchers exploring innovative ADCs therapies, igniting in-depth contemplation for this revolutionary therapeutic strategy.

Breast cancer is one of the prevalent malignant tumors among women worldwide. Conventional treatment strategies often suffer from systemic toxicity, strong drug resistance, and limited targeting capability. In recent years, with advances in biomaterials and drug delivery technologies, microneedles have emerged as a novel transdermal delivery platform offering advantages such as minimal invasiveness, precise targeting, and efficient delivery. These features render microneedles particularly promising for breast cancer therapy. This review summarizes recent progress in the application of microneedle drug delivery systems in breast cancer, including their use in delivering antitumor agents, immunotherapy, photothermal/photodynamic therapy, among others. The mechanisms of action and therapeutic advantages are discussed, along with current challenges and future prospects, aiming to provide a theoretical basis for the translational application of microneedles in breast cancer treatment.

Rheumatoid arthritis (RA), a chronic autoimmune disease, causes progressive cartilage destruction and bone erosion, and also affects vital organs. It affects approximately 0.5%-1% of the global population. Its treatment faces challenges such as insufficient drug targeting, significant systemic side effects and low patient compliance. Responsive microneedle technology integrates inflammatory microenvironment-sensitive materials with intelligent delivery technology to construct a microneedle delivery system that can sense site-specific markers of inflammation (e.g., pH, enzyme activity, or oxidative stress level) to achieve precise local delivery of anti-RA drugs in synovial tissues. Compared with systemic drug delivery, this technology significantly improves local drug concentration in the joints, effectively avoiding the risk of myelosuppression and liver and kidney toxicity associated with traditional therapies. The intelligent response characteristics of microneedles can dynamically regulate the release rate of the drug, so that the drug can be released on demand. The microneedle patch design can significantly reduce the frequency of drug delivery, which significantly improves the long-term treatment adherence. This review systematically explains the classification of responsive microneedles in terms of mechanism of action, material innovation and therapeutic advantages, analyses the current translational bottlenecks, and explores the prospects for their development in the individualized treatment of RA.

The therapeutic efficacy of glioma treatment has long been constrained by the inefficiency of drug delivery due to the blood-brain barrier (BBB). Angiopep-2 peptide actively targets LDL receptor-related protein 1 and efficiently crosses the BBB via LRP1-mediated endocytosis, enabling precise and efficient drug delivery to tumor cells. This offers a novel therapeutic strategy for glioma treatment. Currently, Angiopep-2 has been extensively modified onto various carrier surfaces, significantly enhancing drug accumulation at brain tumor sites. This demonstrates its immense potential in constructing BBB-crossing platforms and has greatly advanced research on brain-targeted delivery systems. This paper primarily reviews the application of Angiopep-2/LRP-1-based modified formulations in glioma treatment. It summarizes research progress across different carriers in glioma, providing a reference for future studies.

The self-assembled nano systems of traditional Chinese medicine (TCM) refer to carrier-free nanostructures spontaneously formed by non-covalent interactions, such as hydrogen bonds, hydrophobic interactions, and π-π stacking, between the active components of TCM. Due to their significant advantages in drug loading, biocompatibility, and intelligent delivery characteristics, these systems have increasingly become a hotspot in the research of TCM modernization, and related research in this area has expanded rapidly. This article reviews the formation mechanisms, advantageous characteristics, and application examples of TCM self-assembled nanostructures from four dimensions: TCM compound formulas, pairs of herbs, single herbs, and individual active ingredients. It delves into the research value of these systems and highlights the key challenges currently faced. The study aims to integrate existing knowledge and provide a theoretical basis for the in-depth exploration and practical application of TCM self-assembly strategies. Furthermore, from the perspective of self-assembly nanotechnology, it not only offers new insights into the material basis of TCM efficacy and the mechanisms of herb combinations but also contributes to advancing TCM research toward precision and modernization.

Peptide therapeutics are characterized by high potency, exquisite specificity, and low toxicity, and have therefore been widely adopted for the clinical management of diverse diseases. However, their inherently low gastrointestinal permeability, rapid enzymatic degradation, and multiple physiological barriers collectively result in extremely low oral bioavailability, necessitating parenteral administration. Compared with the more convenient and patient-compliant oral route, chronic injection is associated with poor adherence and an increased incidence of adverse reactions. In recent years, extensive research has focused on permeation enhancers (PEs) based strategies to improve the oral bioavailability of peptide drugs. This review systematically summarizes the mechanisms of action and clinical applications of representative PEs-including surfactants (sodium caprylate, sodium caprate, alkyl glycosides, bile salts), chelating agents, chitosan and cell-penetrating peptides. It also summarizes related patented technologies (Eligen^TM, TPE^TM, GIPET^TM, POD^TM, Peptelligence^TM) to facilitate the development and clinical translation of oral peptide formulations.

Parkinson's disease (PD) is a common neurodegenerative disorder, characterized by typical symptoms such as resting tremor and muscle rigidity, which can severely impair patients' quality of life. Therefore, early and accurate diagnosis of PD is crucial for improving patient prognosis. Traditional detection methods have limitations including low sensitivity and invasiveness, whereas nanoprobes, with their high specificity, high sensitivity, and multimodal imaging capabilities, offer new opportunities for precise diagnosis and treatment of PD. This article systematically reviews recent advances in the application of nanoprobes for PD detection, with a focus on the classification and functionalization strategies of nanoprobes, as well as their applications in the detection of PD biomarkers and related fields. Additionally, it analyzes current challenges such as biocompatibility and clinical translation, and provides an outlook on future development directions, aiming to offer new insights and references for the early diagnosis and precise treatment of PD.

Trace amine-associated receptor 1 (TAAR1), a widely expressed G protein-coupled receptor in the mammalian central nervous system, serves as a key regulator of monoaminergic and glutamatergic systems and plays a pivotal role in physiological processes such as emotion, cognition, reward, and learning and memory. Recently, research on TAAR1 has expanded beyond its initial focus on psychiatric disorders to rapidly encompass metabolic syndrome and immune system diseases, revealing its considerable potential as a therapeutic target for multiple diseases. This review systematically summarizes TAAR1-associated signaling pathways and their critical roles in various diseases, with a particular emphasis on the therapeutic mechanisms and application prospects of TAAR1 agonists in schizophrenia, depression, Alzheimer's disease, Parkinson's disease, substance use disorder, and type 2 diabetes. Furthermore, this article examines recent advances in the development of TAAR1 agonists, spanning from derivatization tactics based on natural ligands to rational design guided by structural biology, and explores future research objectives, including multi-target drug strategies. The aim of this review is to provide a theoretical foundation and insights to support the development and translational application of TAAR1-targeted therapies.

In recent years, tumors have posed a serious threat to human life and health. Although conventional treatments are widely used, they still have significant limitations. As a result, researchers have increasingly focused on nanotechnology to develop more efficient and precise treatment strategies. Natural nanomaterials, including naturally occurring nanoparticles, polymeric nanomaterials derived from natural products, and self-assembled nanomaterials from natural small molecules, have demonstrated broad prospects in the medical field due to their unique physicochemical properties and excellent biocompatibility. This article reviews the current applications and future potential of several commonly used natural organic nanomaterials in tumor diagnosis and treatment. Looking ahead, natural nanomaterials are expected to not only drive innovation in cancer therapeutics and diagnostics and introduce green sustainable concepts into nanomedicine, but also accelerate clinical translation, offering safer and more effective treatment solutions for patients.

Ulcerative colitis (UC) is an intestinal disease characterized by chronic and recurrent inflammation of the colonic mucosal layer, whose main pathological feature is continuous and diffuse mucosal damage which begins in the rectum and extends towards the proximal colon. G-protein-coupled receptor 21 (GPR21) is an orphan G protein-coupled receptor (GPCR), belonging to class A GPCRs. Previous studies have reported that GPR21 is closely related to diabetes and inflammation. Levistilide A (LA), a natural terpene lactone active component mainly exists in the traditional Chinese medicinal herbs Ligusticum chuanxiong and Angelica sinensis, exhibiting multiple pharmacological activities including anti-inflammation, anti-fibrosis, and anti-tumor. In this study, GPR21-HEK293 recombinant cells were constructed by lentivirus transfection. LA was screened out as a potential GPR21 inhibitor by cell membrane chromatography. RAW264.7 cells were treated with lipopolysaccharide in vitro to establish an inflammatory cell model, which confirmed the correlation between the therapeutic effect of LA on inflammation and GPR21. Dextran sodium (DSS) was used to administrate C57BL/6 mice in vivo to induce a mouse model of UC, at the meanwhile, the mice in LA group were intragastrical administration with LA to investigate its therapeutic effect on UC. In conclusion, we demonstrated that LA as a GPR21 inhibitor improves DSS-induced UC through reducing intestinal inflammation. This work was approved by the Institutional Animals Care and Use Committee of Xi'an Jiaotong University (approval number: XJTUAE2024-1523) and strictly carried out in accordance with their requirements.

Bladder cancer has a high incidence and mortality rate worldwide. Existing treatment methods often face the challenges, such as drug resistance, recurrence and metastasis. These challenges drive our effect to explore new therapeutic compounds. 19-Hydroxycytochalasin B (19-HCB) is a new cytochalasin isolated from marine fungus. Previous studies have confirmed that its structural analogues have good anti-tumor activity, but the anti-tumor activity and mechanism of 19-HCB remain unclear. This study aims to evaluate the in vitro anti-tumor activity of 19-HCB and explore the mechanism of its action in human bladder cancer J82 cells. The anti-tumor activity was comprehensively evaluated through analyzing cell growth by MTT assay, cell scratch assay, high-content imaging and flow cytometry, colony formation and other experiments. The results showed that 19-HCB exhibited the strongest proliferative inhibitory activity (IC₅₀ = 0.59 μmol·L^-1) against bladder cancer J82 cells and could significantly inhibit the migration of J82 cells. Its mechanism of action is closely related to inducing apoptosis, manifested as the upregulation of pro-apoptotic proteins BIM and BAX and the down-regulation of anti-apoptotic protein survivin. Meanwhile, the expression of cell cycle regulatory protein p21 is upregulated while that of CDK1 is downregulated. Additionally, 19-HCB can also inhibit the AKT signaling pathway. In summary, 19-HCB exerts anti-bladder cancer effects by inducing apoptosis, inhibiting migration and regulating multiple key signaling pathways, and is a candidate drug lead compound with development potential.

Serum metabolomics reveals the mechanism of Suanzaoren Decoction (SZRD) in ameliorating anxiety-like behaviors in rats. Insomnia was induced by the modified multiple platform method. Compared with the control group, rats subjected to 18-hour daily sleep deprivation for 21 days exhibited significantly increased total locomotor distance and center zone exploration time (P < 0.01), alongside markedly reduced open arm entry time percentage and frequency (P < 0.001), indicating anxiety-like behaviors. SZRD administration significantly decreased locomotor activity and center zone duration (P < 0.01), while increasing open arm entry time and frequency (P < 0.001), thereby alleviating insomnia-associated anxiety. Pathological sections and immunofluorescence results revealed that, compared with the control group, the model group exhibited inflammatory infiltration in the intestinal mucosal epithelial cells, along with significantly decreased mean fluorescence intensities of ZO-1, occludin, and claudin-1 in the colonic tissue, together with increased hepatic lipid accumulation. SZRD ameliorated intestinal and hepatic damage while enhancing tight junction protein expression. GC-MS analysis demonstrated that the significantly reduced levels of acetic acid, propionic acid, isobutyric acid, valeric acid, isovaleric acid, and isocaproic acid in the colonic contents of sleep-deprived rats (P < 0.05) were significantly reversed by SZRD. UPLC-Q-TOF-MS/MS-based serum metabolomics identified 53 differential metabolites in model rats compared to controls; SZRD significantly reversed 36 of these, encompassing amino acids, organic acids, and lipids. KEGG pathway analysis indicated that SZRD primarily modulated phenylalanine, tyrosine and tryptophan biosynthesis, tryptophan metabolism, tyrosine metabolism, and the tricarboxylic acid cycle. These findings demonstrate that SZRD ameliorates intestinal and hepatic inflammatory injury, reduces intestinal permeability, and exerts anti-insomnia and anxiolytic effects primarily through regulating amino acid metabolism and the tricarboxylic acid cycle, providing scientific evidence for elucidating the therapeutic characteristics of SZRD in treating "consumptive disease with restlessness and insomnia". This experiment was approved by the Animal Ethics Committee of Shanxi University of Chinese Medicine (Approval No.: AWE202310391).

This study aimed to investigate the protective effect and potential mechanism of total flavonoids of Dracocephalum moldavica L. (TFDM) against doxorubicin (DOX)-induced cardiotoxicity (DIC) via the STING/NLRP3 signaling pathway. The protocol was approved by the Animal Ethics Committee of Xinjiang Institute of Materia Medica (approval No.: XJIMM-20250201). The DIC model was established by a single intraperitoneal injection of DOX (20 mg·kg^-1). Body weight, heart weight, heart index, and cardiac function (evaluated by echocardiography) were measured in each group. HE staining and Masson staining were used to observe myocardial tissue damage. Myocardial enzyme activities in serum, as well as oxidative stress indicators and inflammatory factor levels in myocardial tissue, were detected by corresponding kits. Western blot was performed to assess the expression of STING/NLRP3 pathway-related and pyroptosis key proteins in myocardial tissue. Additionally, the effects of TFDM (100 µg·mL^-1) on inflammatory factors and the STING/NLRP3 pathway in rat H9c2 cardiomyocytes stimulated by DOX or the STING agonist vadimezan (DMXAA) were further evaluated in vitro. In vivo experiments showed that TFDM effectively increased body weight, heart weight ratio, left ventricular ejection fraction (LVEF), and left ventricular fractional shortening (LVFS), while decreasing left ventricular internal diameter at end-diastole (LVIDd) and left ventricular internal diameter at end-systole (LVIDs) in DIC mice. TFDM inhibited the abnormal elevation of myocardial enzymes, oxidative stress indicators and inflammatory factors, and ameliorated myocardial inflammatory infiltration and fibrosis. Furthermore, TFDM suppressed the activation of the STING/NLRP3 pathway and the expression of pyroptosis-related proteins in DIC mice. In vitro results demonstrated that TFDM intervention reduced the release of inflammatory factors; Western blot confirmed that both DOX and DMXAA significantly upregulated the expression of STING pathway and pyroptosis-related proteins, which were markedly downregulated after TFDM treatment. In conclusion, TFDM pretreatment exerts a protective effect against DIC by alleviating myocardial injury and improving cardiac function, and its mechanism may be associated with the inhibition of the STING/NLRP3 pathway to suppress cell pyroptosis and inflammatory cascade reaction.

Mycobacterial polyketide synthase 13 (Pks13) is responsible for catalyzing the synthesis of the precursor of mycolic acids and represents a promising target for the development of novel antituberculosis drugs. Based on the hit compound 6 previously identified through virtual screening, a series of benzofuran-3(2H)-one derivatives were designed and synthesized using a structure-based drug design strategy. Among them, compounds 7k and 7o exhibited potent inhibitory activities against Pks13, with IC₅₀ values of 1.01 and 0.64 μmol·L^-1, respectively. This study lays a foundation for the investigation of novel Pks13 inhibitors.

Secreted aspartic protease 2 (SAP2) stands as one of the most pivotal virulence factors in Candida albicans (C. albicans). Consequently, the development of potent small-molecule inhibitors targeting SAP2 represents a critical stride toward the treatment of invasive candidiasis. In our previous studies, the lead compound SAP2-IN-1 was successfully identified and synthesized via integrated strategies of virtual screening and scaffold hopping. Preliminary structure-activity relationship (SAR) investigation was performed through rational design, synthesis, and biological evaluation of 10 novel indolin-2-one derivatives derived from the SAP2-IN-1 scaffold. Enzymatic inhibition and cell-based assays confirmed that compound A1 possesses favorable SAP2 inhibitory activity, with an IC₅₀ of 0.87 μmol·L^-1. Notably, when combined with fluconazole (FLC), compound A1 effectively suppressed the growth of fluconazole-resistant C. albicans 0304103, yielding fractional inhibitory concentration index (FICI) of 0.047. Furthermore, preliminary mechanistic studies revealed that A1 significantly impeded biofilm formation and hyphal transition in drug-resistant C. albicans. It possesses great potential as a novel antifungal lead compound and provides a new theoretical basis and research strategy for the development of virulence factor-targeted inhibitors to overcome the drug resistance of C. albicans.

This study aims to investigate the chemical constituents of the soft coral Sinularia sp. collected from Ximao Island in the South China Sea, and to evaluate the peroxisome proliferator-activated receptor (PPARs) agonistic and anti-inflammatory activities of the isolated compounds. The chemical constituents were isolated using silica gel column chromatography, Sephadex LH-20 gel column chromatography, and high-performance liquid chromatography (HPLC). The structures of the compounds were elucidated by comprehensive analysis of NMR, MS, and IR data and comparing them with those reported in the literature. Eleven compounds were isolated from the soft coral Sinularia sp., including two norsesquiterpenes, two furanosesquiterpenes, and seven steroids. Their structures were elucidated by spectroscopic data analysis as (2E,6E)-3,7-dimethyl-nona-2,6,8-trienoic acid (1), (3S,6E)-3,7-dimethyl-nona-6,8-dienoic acid (2), (1′E,5′E)-5-(2′,6′-dimethylocta-1′,5′,7′-trienyl)-furan-3-carboxylic acid (3), (5′E)-5-(2′,6′-dimethylocta-5′,7′-dienyl)-furan-3-carboxylic acid (4), (24S)-24-methylcholesterol (5), 24R-7α- hydroxyergosta-5-en-3β-ol (6), brassicasterol (7), pregn-5,20-dien-3β-ol (8), gorgosterol (9), β-sitosterol (10), 24-methylenecholesterol (11). Compound 1 was identified as a new compound. Compounds 6 and 10 were obtained from Sinularia sp. for the first time. Luciferase reporter gene assay revealed that compounds 8 and 9 selectively activated the PPAR-α transcription factor at a concentration of 5 μmol·L^-1, while compound 5 acted as a dual agonist of PPAR-α and PPAR-β isomers. In addition, compounds 5, 7, and 8 exhibited in vitro anti-inflammatory activities. Compound 5 significantly inhibited the production of the inflammatory mediators inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS) stimulated mouse macrophages RAW264.7.

Thirteen oligosaccharides were isolated from the water-eluted fraction of Scutellaria baicalensis Georgi extract on macroporous adsorption resin, using techniques such as medium-pressure liquid chromatography, semi-preparative high performance liquid chromatography (HPLC), and a HPLC system equipped with a charged aerosol detector (CAD) as a trigger for fraction collector automatically. The structures of these compounds were identified by mass spectrometry (MS) and nuclear magnetic resonance spectroscopy (NMR) as α-D-galactopyranosyl-(1→6)-α-D-glucopyranosyl-(1→2)-β-D-fructofuranosyl-(6→1)-α-D-galactopyranosyl-(6→1)-α-D-galactopyranoside (1), α-D-galactopyranosyl-(1→6)-α-D-galactopyranosyl-(1→6)-α-D-glucopyranosyl-(1→2)-β-D-fructofuranosyl-(4→1)-α-D-galactopyranoside (2), and α-D-galactopyranosyl-(1→2)-α-D-galactopyranosyl-(1→6)-α-D-galactopyranosyl-(1→6)-α-D-glucopyranosyl-(1→2)-β-D-fructofuranoside (3), raffinose (4), stachyose (5), verbascose (6), ajugose (7), umbelliferose (8), erlose (9), loliose (10), α-D-galactopyranosyl-(1→6)-α-D-galactopyranosyl-(1→6)-α-D-glucopyranosyl-(1→2)-β-D-fructofuranosyl-(1→1)-α-D-galactopyranoside (11), rehmaglupentasaccharide E (12), and rehmaglupentasaccharide F (13). All oligosaccharides with degrees of polymerization ranging from 3 to 6 were isolated from Scutellaria baicalensis Georgi for the first time. Compounds 1-3 are new compounds belonging to the non-classical raffinose-family oligosaccharides. Furthermore, compounds 11-13 are non-classical raffinose-family oligosaccharides and compounds 4-7 are raffinose-family oligosaccharides.

Using various separation and purification techniques including silica gel and recrystallization, two new cyclic dipeptides involving non-proteinogenic amino acids were first isolated and purified from the ethyl acetate fraction of the ethanol (95%) extract of Angelica dahurica. Through the modern spectroscopic methods such as NMR, HR-ESI-MS, IR, OR, UV, and CD, their structures were identified as angelidaphine A (1) and angelidaphine B (2).

A total of four alkaloid compounds, serratinine A (1), fawcettimine (2), 7-hydroxylycopodine (3), and huperzine I (4) were isolated from Huperzia serrata by AB-8 macroporous adsorption resin, Octadecylsilyl, Sephadex LH-20 and silica gel column chromatography combined with the semi-pre HPLC. The isolated compounds were structurally characterized by mass spectrometry (MS), nuclear magnetic resonance spectroscopy (NMR), X-ray single crystal diffraction and circular dichroism (CD). Among them, compound 1 was a square crystalline form with the stereo configuration of (4R, 7S, 8S, 12S, 13S, 15S)-serratinine, named serratinine A. Compound 1 is a new serratinine alkaloid, the first crystal culture and stereo configuration of compound 1 were confirmed in this study.

Hepatic injury caused by pyrrolizidine alkaloids (PAs) has attracted widespread global attention. Establishing a rapid evaluation model for PA toxicity is of great significance for risk assessment of PA exposure. Therefore, this study used senecionine as a representative component to investigate its metabolic characteristics in adult zebrafish, providing core metabolic support for the use of the zebrafish model in rapid screening and evaluation of PA toxicity. Adult zebrafish were orally gavaged with senecionine (0.6 mmol·kg^-1). Liver tissues were collected at 0.25, 0.5, 1, 2, 6, 12 and 24 hours post-administration. Metabolites were identified using ultra-performance liquid chromatography coupled with tandem mass spectrometry (LC-MSⁿ). Concentration-time profiles of major metabolites were determined using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), and pharmacokinetic parameters were fitted. Five metabolites of senecionine were identified in the zebrafish liver, including N-oxide, hydroxylation products, N-glucuronide conjugate of senecionine, and glucuronide conjugate of senecionine hydroxylation product. The parent compound and phase Ⅰ metabolites reached their peaks and were cleared rapidly. Among the phase Ⅱ metabolites, the glucuronide conjugate of the hydroxylation product showed the latest peak and became the predominant form in the zebrafish liver at later time points. This study for the first time systematically elucidated the metabolic profile and kinetic patterns of senecionine in adult zebrafish, confirming that zebrafish possess glucuronidation detoxification capabilities similar to humans. These findings provide a crucial experimental basis for using zebrafish as a model for PA metabolism and toxicity studies, and also offer scientific data for the risk assessment of traditional Chinese medicine preparations containing PAs. This experiment was approved by the Laboratory Animal Ethics Committee of Zhejiang University (Grant No: ZJU20250602).

To identify a new treatment strategy for acute severe pneumonia and solve the bioavailability of apigenin in formulation, the research utilized dipalmitoylphosphatidylcholine (DPPC), as well as 2-hydroxypropyl-β-cyclodextrin as respiratory carriers, to develop the apigenin β-cyclodextrin inclusion complexe DPPC liposomal dry powder inhalers (ACDLDPIs). The ACDLDPIs were prepared by grinding, film-dispersion and freeze drying. The formulation was optimized by orthogonal design, and its particle size, entrapment efficiency, electric potential, in vitro release and lung deposition were studied. Lipopolysaccharide (LPS)-induced rat acute lung injury model was used to evaluate acute severe pneumonia in vivo. The optimal formulation for apigenin cyclodextrin inclusion was obtained with the mole ratio of apigenin/cyclodextrin as (2∶1), concentration of cyclodextrin as 20% and grinding time as 90 min. The optimized prescription of the ACDLDPIs was apigenin inclusion complex/DPPC (1∶2), 15 min hydration time and the mass ratio of mannitol/DPPC (1∶1). Its encapsulation efficiency was (71.46 ± 3.18)%, and the drug loading was (8.36 ± 0.42)%. The particle size was 198.6 ± 4.4 nm (before rehydration) and 221.6 ± 5.5 nm (after rehydration). The zeta potential was 15.3 ± 2.1 mV. The aerodynamic particle size was 4.28 ± 0.77 μm. The pulmonary deposition rate (FPF) was 43.02%. The experiment was approved by the Experimental Animal Ethics Committee of Tongji University Affiliated Yangpu Hospital (No: LL-2023-ZRKX-002). The pharmacodynamic evaluation of acute severe pneumonia was evaluated in vivo. ACDLDPIs group had better therapeutic effect than apigenin group. The ACDLDPIs had suitable particle size, uniform distribution, good stability, high lung deposition rate and good therapeutic effect on acute severe pneumonia in vivo. The ACDLDPIs are promising medicines for the treatment of acute severe pneumonia.

This study utilized monomethoxyl poly (ethylene glycol) 2000-poly (ε-caprolactone) 2000 (mPEG_2K-PCL_2K) and a self-synthesized functional carrier material, fructose-polyethylene glycol 2000-deoxycholic acid (Fru-PEG_2K-DC), which targets the apical sodium-dependent bile acid transporter (ASBT) and glucose transporter 5 (GLUT5) on intestinal epithelial cells, to construct fructose-polyethylene glycol 2000-deoxycholic acid-polymeric micelles/curcumin (FD-PMs/Cur) with the aim of improving drug transintestinal transport efficiency. First, Fru-PEG_2K-DC was synthesized via an azide-alkyne click chemical reaction, and the structure of the synthesized product was confirmed by nuclear magnetic resonance (NMR) and infrared spectroscopy (IR). Subsequently, the film dispersion method was employed to prepare curcumin-loaded targeted micelles. The preparation process and formulation of the drug-loaded nanomicelles were optimized and screened based on evaluation indicators such as particle size, polymer dispersity index (PDI), encapsulation efficiency, and drug loading capacity. Finally, in vitro cell experiments were conducted to evaluate the cellular uptake of the constructed targeted formulation and lysosomal escape function. The prepared FD-PMs/Cur exhibited a near-spherical shape, uniform size, and good dispersibility, with an average particle size of 25.86 ± 0.160 4 nm, PDI of 0.169 ± 0.015, and surface charge of -5.74 ± 0.707 mV. The encapsulation efficiency and drug loading capacity were (92.31 ± 0.40)% and (7.69 ± 0.04)%, respectively. The micelles demonstrated slow release and high stability in gastrointestinal fluids. In vitro cell experiments revealed that the targeted micelles significantly enhanced cellular uptake compared to free drugs and non-targeted micelles, indicating excellent intestinal targeting ability and lysosomal escape function. The constructed FD-PMs/Cur exhibited good gastrointestinal stability, intestinal epithelial cell-targeting capability and lysosomal escape function, effectively enhancing transintestinal transport efficiency. This study provides a novel strategy for improving the oral bioavailability of lipophilic drugs.

This study preliminarily explored the application of gastrodin (GAS) as a brain-targeting ligand for the construction of a novel glucose transporter (glucose transporter type 1, GLUT1) mediated brain-targeted lipid drug delivery system, aiming to achieve cascade-enhanced effects across the blood brain barrier and subsequent tumor targeting. Gastrodin-modified liposomes were successfully prepared using the thin-film hydration method, and their physicochemical properties, drug release characteristics, as well as in vitro and in vivo targeting abilities were preliminarily evaluated. Particular attention was paid to the in vitro and in vivo performance of gastrodin-modified liposomes prepared via a tri-branched conjugation strategy (GAS₃-Lip). The gastrodin-modified liposomes exhibited favorable targeting to bEnd.3 brain microvascular endothelial cells and GL261 glioma cells, as well as significant accumulation at the lesion site in vivo. In both in vitro and in vivo pharmacodynamic evaluations, the tri-branched gastrodin-modified doxorubicin-loaded liposomes (GAS₃-Lip/DOX) demonstrated stronger inhibitory effects against glioma compared with the classic glutathione (glutathione, GSH) modified brain-targeting liposomes, and the relevant animal experiments were approved by the Animal Experiment Ethics Committee of Southern Medical University (No: SMUL202412013). In conclusion, this strategy may provide a valuable approach for drug delivery research targeting glioma therapy.

This study developed a β-asarone (β-As)-modified nano delivery system loaded with tanshinone IIA (As-TSA-NPs) to enhance its therapeutic efficacy against cerebral ischemia-reperfusion injury (CIRI) via intranasal administration. As-TSA-NPs were prepared using the double emulsion-solvent evaporation method and characterized for morphology, particle size, zeta potential, drug loading capacity, and in vitro release behavior. Using coumarin 6 as a fluorescent probe, the 16HBE cell model was employed to investigate the effect of β-As modification on nanoparticle nasal mucosal permeability and inhibition of P-glycoprotein (P-gp) efflux. The therapeutic efficacy was evaluated in vivo using a rat middle cerebral artery occlusion (MCAO) model by assessing neurological function scores, cerebral infarction volume, and oxidative stress markers [superoxide dismutase (SOD), malondialdehyde (MDA)]. Results demonstrated that As-TSA-NPs were spherical, with an average size of approximately 180 nm, a zeta potential of about -33 mV, a drug loading capacity of approximately 3.5%, and exhibited favorable sustained release properties. β-As modification significantly enhanced cellular uptake (1.8-fold higher than unmodified NPs) and improved nasal mucosal permeability by inhibiting P-gp efflux. In vivo studies confirmed that the As-TSA-NPs group showed a 13.5% reduction in cerebral infarction volume, a 13.6% increase in SOD activity, a 21.5% decrease in MDA levels, and significantly improved neurological function scores compared to the model group. This study indicates that β-As-modified nanoparticles effectively enhance the nose-to-brain delivery efficiency of TSA, providing a novel targeted delivery strategy for CIRI treatment. All animal procedures and manipulations in this study were approved by the Animal Ethics Committee of Shanghai University of Traditional Chinese Medicine (PZSHUTCM210226002) and conducted in accordance with its guidelines.

Pulmonary hypertension (PH) is a fatal disease characterized by pulmonary vascular remodeling, in which pulmonary arterial hypertension (PAH) requires long-term combination drug therapy. In this study, we developed bosentan (BST) and sildenafil citrate (SIL) combination tablets to address the complexity of clinical combination drug regimens and poor patient compliance. An HPLC method for the determination of the content at pH 1.2, 4.5 and 6.8 was established through pre-prescription studies, which confirmed that the solubility of BST increased with the increase of pH, while the opposite was true for SIL. The prescription was optimized using wet granulation process with dissolution and content uniformity as the core indexes, and the optimal composition was determined as 47.8% BST, 31.2% SIL, 8%-18% maize starch-pre-gelatinized starch (2∶1), 2%-8% sodium carboxymethyl starch, 3% behenic acid glycerol ester and 1% magnesium stearate. Quality evaluation showed that the tablets complied with pharmacopoeial standards and had good stability. Pharmacokinetic studies in SD rats showed that the t_max of BST and SIL in the compounded tablets were 3.76 and 0.94 h, respectively, which were comparable to those of the commercially available preparations and the half-life was extended by 3-fold and 2-fold, respectively. This compound preparation significantly improves patient compliance by simplifying the dosing regimen (43 mg BST+28 mg SIL, 3 times daily). The two drugs can reach blood drug concentration at the same time, creating favorable pharmacokinetic conditions for achieving synergistic effects at the pharmacological level. This characteristic makes the compound preparation a potential new strategy for the treatment of PAH. The study was approved by the Animal Ethics Committee of China Pharmaceutical University, and all experiments complied with ethical regulations (Approval number: YSL-202505069).

In this study, paeoniflorin (PF), the active constituent of Paeonia lactiflora, was encapsulated within Paeonia lactiflora derived extracellular vesicle-like particles (PL-EVLP) to augment anti-skin-aging efficacy. A hydrogel system (PF@PL-EVLP Gel) was subsequently engineered to further enhance the formulation's stability and transdermal delivery efficiency. PL-EVLP were isolated by differential centrifugation, and paeoniflorin-loaded PL-EVLP (PF@PL-EVLP) were prepared via ultrasound-assisted freeze-thaw cycling. The resulting PF@PL-EVLP exhibited cup-shaped, bilayer-membrane nanostructures with a mean diameter of approximately 148.07 nm, a high drug-loading capacity, and favorable stability. PF@PL-EVLP Gel was fabricated using Carbomer 934 as the gel matrix. Within the hydrogel system, extracellular vesicles maintained structural integrity and uniform dispersion, exhibiting excellent stability. In vitro release studies demonstrated that both PF@PL-EVLP and PF@PL-EVLP Gel exhibited sustained-release characteristics. In vitro pharmacodynamic studies revealed that PF@PL-EVLP exerts robust protection against oxidative damage, significantly elevating intracellular superoxide dismutase (SOD) and hydroxyproline (HYP) levels while markedly reducing malondialdehyde (MDA), matrix metalloproteinase-1 (MMP-1), and reactive oxygen species (ROS) contents. Compared with the solution group, PF@PL-EVLP Gel increased the cumulative transdermal permeation by approximately 10.82-fold, the local skin retention by 7.38-fold, and the cumulative permeation rate by 9.36-fold. In vivo pharmacodynamic evaluations demonstrated that PF@PL-EVLP Gel exerts pronounced anti-skin-aging effects, as evidenced by increased skin hydration, elevated dermal collagen-fiber density, and prevention of D-galactose-induced epidermal and dermal thinning. ELISA analyses revealed that the hydrogel significantly augmented cutaneous total antioxidant capacity (T-AOC), catalase (CAT), SOD, HYP, and tissue inhibitor of metalloproteinase-1 (TIMP-1) levels, concomitantly attenuating dermal MDA and MMP-1 contents; serum SOD was also upregulated and MDA downregulated. Western blot confirmed that the formulation mitigates keratinocyte apoptosis via BAX down-regulation. These findings provide a robust foundation for the topical application of Paeonia lactiflora extracellular vesicle-based hydrogel systems in transdermal delivery of active phytoconstituents for skin-aging retardation. The animal experiments were approved by the Animal Ethics Committee of Shanghai University of Traditional Chinese Medicine (approval numbers: PZSHUTCM2503120002, PZSHUTCM230907004).

Accurate authentication of components in traditional Chinese patent medicines (TCPMs) is fundamental to ensuring the safety and accuracy. Analysis of whole-GEnome (AGE) has been applied to species identification across plants, animals, and fungi. However, its applicability to the authentication of components in TCPMs remains to be evaluated. In this study, AGE was performed to identify Panax notoginseng in TCPMs. Bioinformatic analysis of nine genome assemblies from six Panax species yielded 4 289 410 specific target sequences of P. notoginseng, from which a species-specific target sequence database was constructed. Target sequences were selected and validated by Sanger sequencing. Four molecular techniques including PCR combined with agarose gel electrophoresis, PCR coupled with Sanger sequencing, TaqMan probe-based quantitative PCR, and high-resolution melting analysis were employed to detect the target sequence, thereby establishing identification methods for P. notoginseng. These assays were further applied to commercial TCPMs, all of which were successfully detected to contain P. notoginseng. Therefore, AGE enables accurate identification of components in TCPMs, offering new insights and technologies for quality control of TCPMs.

Steroidal saponins are major active constituents of Maidong (Ophiopogon japonicus (L. f.) Ker Gawl.), and their structures and pharmacological activities are strongly influenced by glycosylation, whereas glycosyltransferases from Maidong remain poorly studied. In this study, we performed de novo transcriptome sequencing, assembly and annotation of roots, stems and leaves of O. japonicus, and combined domain-based screening to identify and conduct phylogenetic analysis of the UDP-dependent glycosyltransferase (UGT) gene family. In total, we obtained 109 UGT candidate coding sequences (CDSs) and identified 34 full-length UGT CDSs (OjUGT1-OjUGT34). Phylogenetic analysis grouped OjUGTs together with Arabidopsis UGTs into 15 subfamilies, among which clade D (UGT73 subfamily) and clade G (UGT85 subfamily) contained relatively more members. Conserved motif analysis and multiple sequence alignment showed that 31 OjUGT proteins possess the canonical plant secondary product glycosyltransferase (PSPG) motif. Expression profiling revealed pronounced tissue-specific differences in OjUGT transcripts, with OjUGT32 (UGT73 subfamily) highly expressed in tuberous roots and OjUGT34 (UGT73 subfamily) highly expressed in leaves. Comparative analyses across species suggested lineage-specific expansion of UGT73 and UGT85 genes within Asparagaceae, which may be associated with steroidal saponin biosynthesis. By integrating phylogenetic relationships, PSPG motif conservation, three-dimensional structure prediction and tissue-specific expression patterns, we propose that OjUGT32 and OjUGT34 are likely involved in the glycosylation of steroidal saponins in O. japonicus. Overall, this study establishes the expression profile of the UGT gene family in Maidong and predicts the physicochemical properties and sequence features of OjUGTs, providing candidate genes for elucidating the glycosylation steps in steroidal saponin biosynthesis.

(-)-Germacrene D is a monocyclic sesquiterpenoid with significant biological activity, and serves as a key precursor in the synthesis of other bicyclic sesquiterpene skeletons. Although plant-derived germacrene D synthases have been widely reported, fungal germacrene D synthases, particularly from genus Penicillium, remain rarely documented. In this study, a sesquiterpene synthase gene, PhGDS, was cloned from the endophytic fungus Penicillium herquei of Huperzia serrata. The encoded protein contains the canonical DDXXD and NSE/DTE motifs of type Ⅰ terpene synthases. To elucidate its function, PhGDS was heterologously expressed in Saccharomyces cerevisiae and Escherichia coli, the resulting products were identified by gas chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance spectroscopy. Two compounds were isolated from the fermentation products of S. cerevisiae, which were identified as (-)-germacrene D (66.01 mg·L^-1) and germacra-1(10),5-dien-4α-ol (0.54 mg·L^-1). Soluble expression of PhGDS was successfully achieved in E. coli, and in vitro enzymatic assays demonstrated that it catalyzes farnesyl diphosphate (FPP) to produce (-)-germacrene D and germacra-1(10),5-dien-4α-ol. This study reports the discovery and characterization of a germacrene D synthase from genus Penicillium, providing a valuable genetic resource for the scalable production of (-)-germacrene D and its derived sesquiterpenoids via synthetic biology.

The MYC transcription factor is a core regulator in the jasmonic acid signaling pathway, widely involved in plant growth and development, secondary metabolism, stress response, and hormone signal transduction. Based on the genomic data of Lonicera japonica, this study identified members of the LjMYC gene family through bioinformatics methods, and analyzed their physicochemical properties, evolutionary relationships, conserved domains and gene structures, as well as promoter cis-acting elements. The expression patterns of this gene family in different tissues, different flowering stages, and under methyl jasmonate (MeJA) treatment were also investigated. The results showed that a total of 25 LjMYC genes were identified in Lonicera japonica, distributed across eight chromosomes. Subcellular localization prediction indicated that 22 LjMYCs proteins were localized in the nucleus, LjMYC13 and LjMYC14 in chloroplasts, and LjMYC4 in the cytoplasm. Cis-acting element analysis revealed that the promoter regions of the LjMYCs gene contain elements responsive to light, growth and development, hormones, and stress. RT-qPCR analysis demonstrated that eight genes, including LjMYC8, exhibited tissue-specific expression in floral organs. The expression trends of ten genes, including LjMYC1, were generally consistent during the flowering process (S1-S6): Expression was slightly lower at S2 than at S1, increased at S3, slightly decreased at S4, and remained high at S5 and S6. Exogenous MeJA treatment promoted flowering, whereas delayed flowering treated with sodium diethyldithiocarbamate trihydrate (DIECA). Except for LjMYC20, MeJA treatment induced the expression of nine members, including LjMYC1, while DIECA treatment suppressed their expression. This study systematically reveals the fundamental characteristics of the LjMYC gene family and its expression patterns in different tissues, flowering stages, and in response to jasmonic acid signaling, providing key insights for exploring the functions of this gene family in jasmonic acid signal transduction in Lonicera japonica.

Drought is a critical stress factor severely restricting the growth and development of Thesium chinense Turcz., while the WOX (WUSCHEL-related homeobox) transcription factor family plays an important role in plant growth, development, and stress response. To explore the drought resistance mechanisms of T. chinense and identify relevant WOX genes, this study simulated drought stress using 20% PEG6000, combined with physiological and transcriptomic analyses to reveal its response strategies. The results indicated that drought stress significantly inhibited plant growth and photosynthesis but activated the antioxidant enzyme system (with CAT, POD, SOD, and APX activities reaching 4.09, 1.57, 3.05, and 58.44 times those of the control, respectively). Additionally, it promoted the accumulation of secondary metabolites such as total polysaccharides, total flavonoids, and thesiumine Ⅰ. Through transcriptomic analysis, the candidate gene TcWOX8 was identified. TcWOX8 was successfully cloned from T. chinense, with a full length of 897 bp encoding 298 amino acids, a protein molecular mass of 33.6 kD, and containing a conserved Homeodomain. Phylogenetic analysis revealed its closest relationship to WOX8 from Vitis vinifera and Hordeum vulgare, suggesting its potential involvement in plant abiotic stress responses. Further experiments confirmed that the TcWOX8 protein can be solubly expressed, possesses transcriptional activation activity, and localizes to the nucleus. The expression of this gene was not only induced by drought stress but also significantly responded to salt, low-temperature, and high-temperature stresses. These results provide a solid molecular foundation for elucidating the drought resistance mechanisms of T. chinense and revealing the key regulatory role of TcWOX8 in broad-spectrum stress defense in plants.